Fig. 3: Characterisation of nerolidol-producing strains, harbouring nerolidol synthetic genes on a 2μ plasmid (N401-1) or integrated at amplified RPL25 locus (N401-2, N401-3, and N401-4).
From: An in vivo gene amplification system for high level expression in Saccharomyces cerevisiae
a, b Schematic map of genetic vectors used to introduce nerolidol synthetic genes into yeast. c–h Strain characterisation in two-phase flask cultivation with 20 g l−1 glucose and dodecane overlay. Y-FAST (fluorescence-activating and absorption-shifting tag) fluorescence was measured after 4-hydroxy-3-methylbenzylidene rhodanine (HMBR) with final concentration 20 μM was added to the yeast samples before flow cytometry assay, and is expressed as fold-change of exponential-phase auto-fluorescence of the reference strain GH43. Nerolidol production at 72 h was shown. Kernel density was calculated with bandwidth equal to 0.05. Mean values ± standard deviations are shown (c–f, h; N = 4 independent biological replicates). Two-tailed Welch’s t-test was used for comparing two groups, and p values were shown in d, h. Source data are provided as a Source Data file.
